Indian Journal of Microbiology Research

6-10

Authors: Jegan C,Mangayarkarasi V,Rukadikar AR*,Anandi V

Introduction: Bacteraemia means presence of microorganisms in circulating blood or blood stream. Septicaemia means multiplication of microorganisms and production of toxins in blood stream. Bacteraemia may be transient, intermittent or continuous. The two categories of blood stream infections are intra vascular and extra vascular. Early microbiological diagnosis and determination of antimicrobial sensitivity pattern have been shown to improve treatment outcome. Aim: To isolate the causative agents and determine the antimicrobial sensitivity pattern from the blood sample of patients with bacteraemia/septicaemia. Materials and Methods: Hospital based prospective cross sectional study conducted at SRM Medical College Hospital and Research Centre, Potheri, Kancheepuram District. Total 765 blood sample were collected during March 2012 to February 2013 from bacteraemia diagnosed cases. The blood samples were collected and processed by standard methods. Isolation and identification of organisms was done as per standard guidelines. Antimicrobial sensitivity was determined by Kirby Bauer’s Disc diffusion method as per CLSI guidelines. Observation and Result: Among 765, 416 (54.37%) were male and 349 (45.62%) were female. Among the total sample of 765, 114 (14.9%) showed positive. Present study showed predominance of Gram positive organisms i.e. 60 (52.66%) over Gram negative organisms, i.e. 51 (44.73%) and 3 (2.63%) Fungal isolates. Conclusion: Timely detection and appropriate treatment will help to reduce the morbidity and mortality of patients. Also, such studies will helps in developing guidelines and antibiotic policy for better outcome of the patients.

15-17

Authors: Sharlee R*,Sumangala B

Introduction: Urinary tract infection (UTI) is an important cause of illness in humans with emergence of multidrug resistance. Fosfomycin remains to be effective against MDR uropathogens. We evaluated susceptibility of Fosfomycin against uropathoges. Materials and Methods: A prospective study was conducted for a period of 6 months in a tertiary care hospital MIMS, Mandya. Significant bacterial growth were identified in urine samples and processed accordingly. Fosfomycin susceptibility was performed by Kirby-Bauer disc diffusion method along with other drugs advised as per CLSI guidelines. Results: A total of 893 urine samples were received, 150 samples showed significant growth of one or two organisms, yielding a sum of 162 isolates. Out of 162 isolates with significant growth of organisms, majority were Gram negative bacilli (GNB) accounting for 63.6% (103 isolates) followed by Gram positive cocci (GPC) 36.4% (59 isolates). Out of 103 GNB, ESBL producer were 25.2%, Carbapenamase producer were 1.9%, Amp C were 6.8%, ESBL+ Carbapenamase were 4.9%, Amp C + Carbapenemase were 2.9% and 58.3% were non ESBL, Carbapenamase and AmpC producer. Out of 162 isolates, 156(96.3 %) were sensitive to Fosfomycin and 6 (3.7%) were resistant to Fosfomycin. Conclusions: Fosfomycin is effective against MDR, ESBL, AmpC and Carbapenamse producing uropthaogens. It is an effective, convenient and safe drug for treating the patients of all age group and pregnant mothers with uncomplicated UTIs.

22-29

Authors: Nirbhavane H M,Bagde U S

The aim of this study was to understand the mechanism of resistance in Enterobacter spp. towards cobalt toxicity. The MIC (minimum inhibitory concentration) and cobalt tolerance determination was done. The cellular biochemistry of this microorganism was checked with the help of estimations of macromolecules, dehydrogenases activity, and determination of cobalt through cell fractionation. Cobalt inhibited the growth and viability of sensitive organisms. Addition of cobalt affected the growth curve phases, which was ultimately an impact of concentration of cobalt. The minimum inhibitory concentration (MIC) determined was 250 ppm in sensitive Enterobacter spp. that completely inhibited the growth, synthesis of protein, DNA, and activity of dehydrogenases of the TCA cycle. The resistant Enterobacter spp. tolerated even 500 ppm concentration of cobalt. The cobalt incorporated in to the cell wall, membrane and cytoplasm was 66%, 11% and 23% and 9%, 58% and 33% in resistant Enterobacter spp. and sensitive Enterobacter spp. respectively when 250 ppm cobalt was added in the culture flasks. The mechanism of resistance was accumulation of cobalt in the cell wall and presence of plasmid.

35-37

Authors: Ujjwal Gohel,Vicky Gandhi*,Alka Nerurkar

Introduction: Rotavirus infection is a leading cause of acute gastroenteritis in children below 5 years of age. Rotavirus infection is estimated 29% of all diarrheal deaths in children below 5 years of age. It affects almost 95% of children under 5 years of age. Aims and Objectives: The study was used to detect prevalence of rotavirus infection in children below 5 years of age presenting with diarrhea and vomiting in a tertiary care hospital. The study was used to analyze the epidemiology of Rotavirus infection in local area. Materials and Methods: All children suffering from acute diarrhea admitted in pediatric ward were included in the study. Stool samples were tested for detection of Rotavirus antigen by ELISA method. Results: Total 60 stool sample were collected during study period of 2 months. 50 samples were collected from symptomatic patients and 10 samples were collected as control samples. Rotavirus antigen was detected from 6 stool samples. Conclusion: Rotavirus infection prevalence is 12% in our study which is low when compared to other studies. The factors attributing to this incidence are seasonal variation, socio-economic status and personal hygiene and sanitation practices. Limitations: As the study duration was 2 months (STS- ICMR Study), further more analysis is required with good sample strength in all seasons to know the epidemiological trends of rotavirus.

44-49

Authors: Sadia Zia,Muhammad Danish Mehmood*,Huma Anwar,Saba Gul,Ejaz Rafique

Primary prerequisite of effective influenza virus immunization is homology of vaccinal strain to the wild type and appropriate immunogen count in each dose of vaccine to induce protective immune response. The purpose of current project is to study the effect of immunogen potency in inactivated monovalent and bivalent avian influenza vaccines. Eight AIV (Monovalent & Bivalent) oil based vaccines with different immunogen level were prepared and evaluated for potency in AIV susceptible broilers through Haemagglutination Inhibition (HI) test on 18 and 36-day post vaccination. It was observed that 0.3ml dose of inactivated avian influenza oil based AIV-H9 vaccine having infectivity titer EID50 1×109.2/ml and (HA= 512) induced high mean anti AIV-H9 antibody titers (72±22.62) as compared to the vaccine containing EID50 1×108.2/ml and (HA= 256). Similarly, same dose of bivalent avian influenza inactivated vaccine containing infectivity titer EID50 1×109.2/ml and (HA= 512) induced better immune response against each antigen H7 and H9 respectively, that of 0.3 ml dose of the vaccine comprising infectivity titer EID50 1×108.2/ml and (HA= 256) where H7 and H9 were documented. It was concluded that infectivity titers in 0.3 ml dose of inactivated adjuvanted vaccine shall be ? 108.2 /ml or 256 HAU to achieve protective anti body titers to get protection against avian influenza virus and its complications over a long period of time thus contributing to disease control in epidemics.

57-60

Authors: Arunava Kali,Kalaivani Ramakrishnan*,Pravin Charles M. V,Sreenivasan Srirangaraj,Seetha K.S

Introduction: Neonatal sepsis is a major cause of neonatal mortality, the clinical outcome of which depends on early diagnosis and initiation of appropriate antibiotics. The emergence of multi-drug resistant strains has limited the choice of available antibiotics. Thus, antibiotic resistance pattern of pathogens iscritical for both therapy and infection control. Objective: To determine the etiological agents of neonatal sepsis and their antibiotic resistance pattern. Materials and Methods: In this study, all neonates with suspected sepsis, admitted to neonatal intensive care unit during January to December, 2014 were included. Aerobic blood culture was done using BACTEC FX system. Identification and antibiotic susceptibility testing of isolates from positive cultures was carried out. Results: Out of 522 neonates who developed clinical sepsis, 64 grew pathogens on blood culture. The most common organisms were coagulase negative Staphylococcus species (CONS)(32.8%), Klebsiella pneumoniae (18.7%), Escherichia coli (9.4%), Enterococcus faecalis (7.8%) and Candida species (7.8%). The gram negative isolates showed high resistance to ampicillin (90%), gentamicin (70%), ceftriaxone (66.7%), cotrimoxazole (56.7%), amikacin (53.3%) and ciprofloxacin (43.3%). In contrast, imipenem (16.7%), piperacillin-tazobactam (26.7%) and cefepime-tazobactam (0%) were effective with lower resistance rates. A large majority of the Enterobactericeae isolates (66.7%) were extended spectrum beta-lactamase producers. Among the gram positive isolates, resistance to penicillin, erythromycin and ciprofloxacin were 93.1%, 62.1%and 51.7% respectively. These strains showed uniform sensitivity to vancomycin, teicoplanin and linezolid. Conclusion: While CONS were the predominant isolate, gram negative bacilli and Enterococcus sp have shown high resistance to commonly used antibiotics. Low resistance was observed with Cefepime-tazobactam, imipenem (for gram-negative isolates) and vancomycin, teicoplanin and linezolid (for gram-positive isolates) in our study.

66-70

Authors: Ramakrishnan Kalaivani,Marie Victor Pravin Charles*,Suresh Sah,Kunigal reenivasiah Seetha

Context: Staphylococcus aureus is a Gram positive bacterium which has an impending ability to cause an array of infection ranging from skin lesions to life threatening systemic illness. In the past few decades there is a drastic rise in Methicillin resistant Staphylococcus aureus (MRSA). The acquisition of Staphylococcal cassette chromosome mec (SCC mec) carrying mecA gene leads to Methicillin resistance among Staphylococcus aureus. Aim: The aim of this study is to identify the prevalence of MRSA and identify the SCC mecA and Panton-Valentine Leukocidin (PVL) gene by Polymerase chain reaction (PCR) among Staphylococcus aureus among various clinical isolates in our hospital. Materials and Methods: A prospective cross-sectional study was conducted in a tertiary care hospital at Puducherry. The study was conducted over a period of 1year from January 2014 to December 2014. The isolates were obtained from various clinical samples such as pus, aspirate, urine, blood and sputum. PCR amplification of mecA and pvl genes was performed to check their prevalence among the isolates collected from a tertiary care hospital. PCR reaction mixtures (10?l) contained 1?lTaq PCR buffer (10X) with 15mM MgCl2, 0.1 µl (0.5U) of Taq polymerase (New England Biolabs, US), 0.25mM of dNTP mix (Thermo Scientific, US), 0.25pM each of forward and reverse primers (Eurofins, Bangalore, India) and 10ng of genomic DNA template. Results: Around 228 samples of Staphylococcus aureus isolated from various clinical samples were analyzed for methicillin sensitivity. Out of these samples around 49 (21.49%) samples were MRSA positive isolates. Out of 49 mecA harboring isolates pvl gene was amplified in 26 (53%) isolates. Conclusion: MRSA being a dreadful pathogen has minimal treatment options. PVL gene with added virulence further worsens the clinical outcome among infected patients. Hence the knowledge of its prevalence adds an insight among the infection control practitioners to adhere effective prevention protocol.

78-81

Authors: Anupama Bahuleyan,Harshan K H*,Geeta Bhai

Introduction: The study was conducted to know about the Gram positive cocci causing urinary tract infections in catheterized patients in intensive care units and to study their antimicrobial sensitivity pattern. Materials and Methods: A sample size of 100 catheterised patients in Intensive care units were included in a period of 1 year from December 2012 to November 2013 to assess the urinary tract infections caused by indwelling catheter. Result: Of 100 cases, 11 showed catheter associated urinary tract infections. Out of them 10 were Enterococcus spp, and 1 was Coagulase negative staphylococci. Isolates were multi drug resistant and showed sensitivity to Vancomycin, Linezolid, Teicoplanin. Conclusion: Infections were more with male sex, prolonged catheterization, old age and diabetes. High incidence of CAUTI was found in the first 2 weeks of catheterization. It was noted by antimicrobial susceptibility, that most of the urinary isolates are resistant to the commonly used antibiotics.

89-91

Authors: Prasanna Nakate,Yogendra Shelke*,Snehal Patil,Suvarna Patil,Vasant Kavade

Introduction: Urinary tract infections (UTI) are one of the common infections during pregnancy. If not treated, these can cause serious complications to both mother and foetus. This study was carried to determine the prevalence of UTI in pregnant women and to identify the common uropathogens causing UTI. Materials and Methods: Midstream urine specimens were collected from 180 pregnant females. The specimens were examined microscopically and by culture. A colony count of 105 CFU/ml of urine of a single uropathogenic bacterium in culture is considered significant and such patients, either symptomatic or asymptomatic, are considered to have a urinary tract infection. Results: Urine specimens of 33 females (18.33%), all of whom were asymptomatic, showed significant growth of uropathogenic organisms confirming the diagnosis of UTI. The commonest pathogen isolated was E. coli (54.55%), followed by Klebsiella species (24.24%), S. aureus (12.12%), Pseudomonas spp, Pseudomonas aeruginosa and C. albicans (3.03% each). Conclusion: In this study, the prevalence of UTI in pregnancy was found to be 18.33%. As most of these cases present without any symptom, its early detection by screening for it with appropriate laboratory tests is vital. This would help clinicians to start appropriate antibiotic treatment at an early stage which in turn will help to minimize complications associated with UTI.

97-100

Authors: M Jane Esther*,P K. Rath,R Arivazhagan

Nocardiae are aerobic gram positive filamentous bacteria that are acid fast when stained by modified acid fast staining. They are called “Aerobic actinomycetes” because of their resemblance to actinomycetes in gram stain. They usually cause infections in immunocompromised individuals but can affect the immunocompetent as well. We have described here the report of an immunocompetent young male who presented with nocardial gluteal abscess. Nocardia must be considered in the differential diagnosis of any systemic illness and must be ruled out prior to making any other diagnosis. Correct suspicion of nocardiosis by the treating physician and the clinical microbiologist and early treatment of nocardiosis can be life saving to a great extent. Gram staining and modified acid fast staining performed together can definitely help in arriving at the exact diagnosis at the earliest.